Synergistic combination of a glyphosate compound and one of dcoit or oit or bbit

ABSTRACT

A synergistic antimicrobial composition containing a glyphosate compound and one of 4,5-dichloro-2-n-octyl-4-isothiazolin-3-one or 2-n-octly-4-isothiazolin-3-one or N-Butyl-1,2-benzisothiazolin-3-one is provided. Also provided is a method of inhibiting the growth of or controlling the growth of microorganisms in a building material by adding such a synergistic antimicrobial composition. Also provided is a coating composition containing such a synergistic antimicrobial composition, and a dry film made from such a coating composition.

This invention relates to combinations of biocides, the combinationshaving unexpectedly greater activity than would be expected for the useof both of the individual antimicrobial compounds.

Use of combinations of at least two antimicrobial compounds can broadenpotential markets, reduce use concentrations and costs, and reducewaste. In some cases, commercial antimicrobial compounds cannot provideeffective control of microorganisms, even at high use concentrations,due to weak activity against certain types of microorganisms, e.g.,those resistant to some antimicrobial compounds. Combinations ofdifferent antimicrobial compounds are sometimes used to provide overallcontrol of microorganisms in a particular end use environment. Forexample, WO 1998/121962 discloses combinations of3-iodo-2-propynyl-butylcarbamate and zinc pyrithione, but this referencedoes not suggest any of the combinations claimed herein. Moreover, thereis a need for additional combinations of antimicrobial compounds withrelatively low impact on health and/or the environment. The problemaddressed by this invention is to provide such additional combinationsof antimicrobial compounds.

Antimicrobial compounds are sometimes included in liquid coatingcompositions that are applied to a substrate and that become dry films.It is desirable that such dry films control surface fungi and algae andthat such dry films also present as little adverse effect as possible onhealth and the environment.

The following is a statement of the invention.

The first aspect of the present invention is a synergistic antimicrobialcomposition containing a glyphosate compound and one of4,5-dichloro-2-n-octyl-4-isothiazolin-3-one or2-n-octly-4-isothiazolin-3-one or N-Butyl-1,2-benzisothiazolin-3-one isprovided. A second aspect of the present invention is a method ofinhibiting the growth of or controlling the growth of microorganisms ina building material by adding the synergistic antimicrobial compositionof the first aspect of the present invention. A third aspect of thepresent invention is a coating composition containing a synergisticantimicrobial composition of the first aspect of the present invention.

A fourth aspect of the present invention is dry film made from a coatingcomposition of the third aspect of the present invention.

The following is a detailed description of the invention.

As used herein, the following terms have the designated definitions,unless the context clearly indicates otherwise.

The term “antimicrobial compound” refers to a compound capable ofinhibiting the growth of or controlling the growth of microorganisms;antimicrobial compounds include bactericides, bacteristats, fungicides,fungistats, algaecides and algistats, depending on the dose levelapplied, system conditions and the level of microbial control desired.The term “microorganism” includes, for example, fungi (such as yeast andmold), bacteria and algae. The following abbreviations are usedthroughout the specification: ppm=parts per million by weight(weight/weight), mL=milliliter, ATCC=American Type Culture Collection,and MIC=minimum inhibitory concentration. Unless otherwise specified,temperatures are in degrees centigrade (° C.), and references topercentages are by weight (wt %). Percentages of antimicrobial compoundsin the composition of this invention are based on the total weight ofactive ingredients in the composition, i.e., the antimicrobial compoundsthemselves, exclusive of any amounts of solvents, carriers, dispersants,stabilizers or other materials which may be present.

As used herein, “DCOIT” is 4,5-dichloro-2-n-octyl-4-isothiazolin-3-one,and “OIT” is 2-n-octly-4-isothiazolin-3-one, and “BBIT” isN-Butyl-1,2-benzisothiazolin-3-one. When a ratio is said herein to be“X:1 or higher,” it is meant that the ratio is Y:1, where Y is X orgreater, and when a ratio is said herein to be “X:1 or lower,” it ismeant that the ratio is Z:1, where Z is X or less.

Glyphosate is N-(phosphonomethyl)glycine (registry number 1071-83-6).Glyphosate is a known herbicide that is approved in many jurisdictionsfor use for weed control on food crop plants. One measure ofglyphosate's relatively low impact on the environment is shown by itsrelatively low activity against various algae. Some results showing theactivity of glyphosate against various algae are as follows:

Test⁽¹⁾ Algae 72 hr 96 hr 7 day E_(b)C₅₀ green algae 485 mg/l 13.8 mg/l(Selenastrum capricornutum) E_(r)C₅₀ green algae 460 mg/l (Selenastrumcapricornutum) EC₅₀ marine algae 1.3 mg/l 0.64 mg/l (Skeletonemacostatum) EC₅₀ diatom (Navicula pelliculosa)   42 mg/l EC₅₀ blue-greenalgae   15 mg/l (Anabaena flos-aquae) ⁽¹⁾Notes: EC_(n)/EC₅₀ EffectiveConcentration EC_(n) is the concentration of a substance that affects n% of a population in a given period of time. EC₅₀ is widely used sinceit is the most accurate point in the concentration effect curve.E_(b)C₅₀ Median effective concentration for biomass (algae): Theconcentration of a substance which reduces by 50% the increase ofbiomass compared with a control sample, over a specified time period.E_(r)C₅₀ Median effective concentration for growth rate (algae): Theconcentration of a substance which reduces by 50% the growth rate ofalgal cells, compared with a control sample, over a specified timeperiod.

The above data show that glyphosate has relatively low impact on algae,which indicates that glyphosate has relatively low impact theenvironment. This characteristic is considered desirable for anantimicrobial material that may be included in a coating composition.

The present invention involves the use of a glyphosate compound. As usedherein “a glyphosate compound” means glyphosate or a glyphosate salt. Aglyphosate salt is a metal salt of glyphosate. Suitable metals includealkali metals, alkaline earth metals, and transition metals. Glyphosatesalts are preferred over glyphosate. More preferred are transition metalsalts of glyphosate; most preferred is zinc glyphosate.

Glyphosate has relatively high solubility in water. This relatively highsolubility in water is not preferred for an antimicrobial material thatmay be included in a coating composition or other building material,because dried coatings and building materials are exposed to water,which could tend to remove a highly soluble compound from the driedcoating or the building material. Preferred are glyphosate salts thathave solubility in water, at 20° C., of 3 g/l or less; more preferred is1 g/l or less; more preferred is 0.3 g/l or less.

It is considered that glyphosate salts will have relatively low impacton health and the environment, because it is considered that glyphosatesalts will have impact on health and the environment that is similar tothe impact of glyphosate.

The present invention involves a composition that contains both aglyphosate compound and one of4,5-dichloro-2-n-octyl-4-isothiazolin-3-one or2-n-octly-4-isothiazolin-3-one or N-Butyl-1,2-benzisothiazolin-3-one. Ithas been surprisingly found that such a composition is synergisticallyeffective as a biocide. It has been especially surprisingly found thatcompositions that contain both zinc glyphosate and one of DCOIT or OITBBIT are synergistically effective as biocides. Preferred arecompositions that contain both a glyphosate compound and DCOIT. It hasbeen even more especially surprisingly found that compositions thatcontain both zinc glyphosate and DCOIT are synergistically effective asbiocides.

When DCOIT is present, preferably the weight ratio of DCOIT toglyphosate compound is 0.33:1 or higher. When DCOIT is present,preferably the weight ratio of DCOIT to glyphosate compound is 2:1 orlower. When OIT is present, preferably the weight ratio of OIT toglyphosate compound is 0.3:1 to 0.4:1. When BBIT is present, preferablythe weight ratio of BBIT to glyphosate compound is from 0.1:1 to 10:1.

If a formulation contains more than one of the listed IT compounds(i.e., DCOIT, OIT, and BBIT), it is useful to consider the sum of theweights of all the listed IT compounds (i.e., the sum of the weight ofDCOIT (if it is present) plus the weight of OIT (if it is present) plusthe weight of BBIT (if it is present). The preferred ratio of the sum ofthe weights of all the listed IT compounds to the weight of glyphosatecompound is 0.1:1 to 10:1.

The mixture of glyphosate compound and one of DCOIT or OIT or BBIT maybe included in a coating composition. Glyphosate compound and one ofDCOIT or OIT or BBIT may be added to the coating composition separatelyor as a mixture or any combination thereof. Preferred coatingcompositions are liquid. Coating compositions may be aqueous ornon-aqueous. Aqueous coating compositions contain 40% or more water byweight, based on the weight of the coating composition.

Among embodiments in which glyphosate compound and one of DCOIT or OITor BBIT are included in a paint or other coating composition, preferredcoating compositions are liquid compositions, especially compositionsthat contain dispersions of polymers in aqueous media.

In addition to paints and other coating compositions, the biocidecombinations of the present invention are particularly useful inpreservation of building materials, e.g., adhesives, caulk, jointcompound, sealant, wallboard, etc., polymers, plastics, synthetic andnatural rubber, paper products, fiberglass sheets, insulation, exteriorinsulating finishing systems, roofing and flooring felts, buildingplasters, bricks, mortar, gypsum board, wood products and wood-plasticcomposites. When a biocide combination of the present invention ispresent in a building material, it is preferred that some or all of thebiocide combination be present at the surface of the building materialor near enough to the surface of the building material to inhibitmicrobial growth on that surface.

In some embodiments, latex paints or other liquid coating compositionsare used that contain the biocide combinations disclosed herein.

Coating compositions are designed so that a layer of the coatingcomposition can readily be applied to a substrate and then dried orallowed to dry to form a dry film. Coating compositions contain abinder. Binders contain one or more of the following: one or morepolymer, one or more oligomer, and/or one or more monomer. Oligomers andmonomers in binders are designed to polymerize and/or crosslink duringor after the formation of the dry film. Polymers in a binder may or maynot be designed to crosslink during or after the formation of the dryfilm.

Coating compositions optionally contain one or more pigment. A pigmentis a mineral or an organic substance in the form of small solidparticles. Pigments provide full or partial opacity to the dry film.

The biocide combinations are useful for preservation of the dry filmcoating resulting after application of a paint or other liquid coatingcomposition. Preferably, the antimicrobial composition is an aqueouslatex paint comprising one or more of the biocide combinations disclosedherein, or the dry film coating resulting from application of the paintto a surface. An aqueous latex paint is an aqueous liquid coatingcomposition in which the binder is an polymer in the form of a latex(i.e., in the form of polymer particles dispersed throughout the water).More preferred are aqueous latex paints in which the binder contains oneor more acrylic polymer.

Typically, the amount of the biocide combinations of the presentinvention to control the growth of microorganisms is from 100 ppm to10,000 ppm active ingredient. Preferably, the active ingredients of thecomposition are present in an amount of at least 300 ppm, preferably atleast 500 ppm, preferably at least 600 ppm, preferably at least 700 ppm.Preferably, the active ingredients of the composition are present in anamount of no more than 8,000 ppm, preferably no more than 6,000 ppm,preferably no more than 5,000 ppm, preferably no more than 4,000 ppm,preferably no more than 3,000 ppm, preferably no more than 2500 ppm,preferably no more than 2,000 ppm, preferably no more than 1,800 ppm,preferably no more than 1,600 ppm. Concentrations mentioned above are ina liquid composition containing the biocide combinations; biocide levelsin the dry film coating will be higher.

The present invention also encompasses a method for preventing microbialgrowth in building materials, especially in dry film coatings, byincorporating any of the claimed biocide combinations into thematerials.

Typically, the antimicrobial compositions are used to inhibit growth ofalgae and/or fungi.

The composition of the present invention contains a glyphosate compoundand one of DCOIT or OIT or BBIT. It is contemplated that someembodiments may contain one or more additional antimicrobial compound.

It is contemplated that zinc glyphosate shows unexpectedly synergisticantimicrobial action in comparison with results that would be expectedfrom a combination of zinc and glyphosic acid.

The following are examples of the present invention.

Zinc Glyphosate was synthesized as follows.

First, pre-preparation was performed as follows. 15-20 g Glyphosate acidwas dried in oven at 80-90° C. overnight. A 1 M solution of NaOH indeionized (DI) water was prepared.

Glyphosate solution was made as follows. 200 mL DI water was measuredinto 600 mL beaker containing stir bar. 15 g Glyphosate acid was slowlyadded to beaker of water while stirring on stir plate at medium speed.Temperature was raised up to 60-70° C. to dissolve the glyphosate;temperature was monitored with thermometer. 1M NaOH was added to bringpH up to 6.0. The glyphosate dissolved at a pH around 2.4. The mixturewas agitated for 5-10 minutes.

A zinc solution was made as follows. 100 mL DI water was measured into400 mL beaker containing stir bar. 36.25 g Zinc Chloride (ReagentGrade, >98%, Sigma-Aldrich, material #208086) was measured into weighboat Zinc chloride was slowly added to water in beaker while mixing onstir plate. Solution was heated up to 65° C.

Zinc glyphosate solution was made as follows. Using a 9-inch Pasteurpipette, the zinc solution was added to the glyphosate solution. Aftereach addition of a small amount of zinc solution to glyphosate solution,the precipitate dissolved before further zinc solution was added. Whenthe precipitate would no longer dissolve, pH was adjusted up to 5.Remainder of zinc solution was poured into the glyphosate solution, andthe mixture was stirred overnight.

NOTE: controlling the pH of the solution is extremely important forobtaining the desired product. During the preparations reported herein,care was taken not exceed pH value of 5.

Zinc Glyphosate Filtration was performed as follows.

Filtration apparatus was set up using a Buchner funnel and a filterflask connected with a flask seal and hooked up to a water-vacuum pump.A #41 Whatman™ filter paper was placed in the funnel. The pump wasturned on, and DI water was poured onto filter paper to create vacuum.The Glyphosate, zinc slurry was slowly poured onto the filter paper andthen washed once with hot (approximately 50° C.) DI water, and thenrinsed twice with isopropyl alcohol (IPA).

The Zinc Glyphosate Final Preparation was performed as follows.

Filter paper containing zinc glyphosate precipitate was removed andplaced in a large Pyrex™ dish. The dish was covered with metal foil, anda few holes were poked in the foil for ventilation. The precipitate wasdried in oven at 80-90° C. overnight. The precipitate was then measuredout into a pre-massed, clean, labeled glass bottle, and the mass of theprecipitate was recorded.

Sample preparation for antimicrobial testing was performed as follows.

A single biocide or blend of biocides was post added into white acryliclatex paint free of biocides to give a maximum total active ingredient/sconcentration tested. This paint was then diluted with a biocide freeacrylic latex paint to give targeted concentrations for the testing.Depending on the type of biocide blends tested, the total biocidesconcentrations varies from 400 to 3300 ppm. After biocides addition ordilution, each sample was hand mixed for at least a minute untiluniformity was achieved. Each of the paint samples as well as a controlsample (containing no biocide) were used to prepare films on blackplastic-vinyl chloride/acetate copolymer panels (Leneta Company, Mahwah,N.J.) using a 0.0762 mm (3 mil) bird bar applicator. The panels werethoroughly dried for at least 2 days avoiding direct exposure tosunlight. Square discs (0.5 inch²; 1.27 cm²) were cut out from eachpanel and were used as the substrate for fungal and algal efficacytests. This sample size allowed for an agar border when the sample discwas placed into the well of the test plate. Each sample was tested induplicate.

The test conditions were as follows.

The appropriate media (BOLD'S 3N for Chlorophytes, BG-11 forCyanobacteria, and PDA for fungi) were used to support microbial growth.The test plates were maintained at room temp (25° C.-26° C.), in acycled light-dark environment, for four weeks for algae. Plates forfungal challenge tests were maintained at 30° C. for four weeks. At theend of the incubation period the samples were scored for percent areacovered by visible microbial growth.

Algal inoculum was as follows.

Medium abbrevi- for Organisms ation Type testing Gloeocapsa sp. Gs ATCC29159 Unicellular, BG-11 Colonial Cyanobacteria Oscillatoria sp. Os ATCC29135 Filamentous BG-11 Cyanobacteria Nostoc commune Nc CCAP 1453/29Unicellular, Bold Cenobial Chlorophyte Trentepohlia Ta + To UTEX LBFilamentous Bold aurea + 429 + Chlorophyte Trentepohlia CCAP 483/4odorata Chlorella sp. Cs + Ck ATCC 30582 + Unicellular Bold UTEX + ATCC11468 Chlorophyte Chlorella kessleri Calothrix parientina Cp UTEX LBFilamentous Bold 1952 Cyanobacteria

Fungal inoculum was as follows.

Medium for Organisms abbreviation ATCC# Growth and Testing Aspergillusniger An 9642 PDA Penicillium funiculosum Pf 11797 PDA Cladosporiumherbarum Ch 11281 PDA Aureobasidium pullulans Ap 9348 PDA Trichodermaviride Tv 32630 PDA Alternaria alternata Aa 20084 PDA

Algal efficacy was tested as follows (Modified ASTM 5589).

ASTM 5589 is a standard accelerated test method for determiningresistance of various coatings (including paints) to algal defacement.To accommodate for high-throughput screening, this method was scaleddown from petri plates to 6-well plates. A single coupon was placed witha pair of sterile forceps at the center of the agar plug (on top) withthe painted surface facing upwards. Algal inoculums were prepared bymixing equal concentrations (approximately 1×10⁶ cfu/ml) and equalvolumes (depending on number of samples to be inoculated) of likegrowing organisms.

In this synergy study, three pool of mixed algae were prepared as thetest inoculum, Gloeocapsa sp. and Oscillatoria sp. a mix ofcyanobacteria grown on BG-11 media; Chlorella sp., Chlorella kessleri,and Nostoc commune are unicellular chlorphytes that were mixed and grownon Bold media; Trentepohlia aurea, Tretepohlia odorata, and Calotrixparientina are filamentous algae that were mixed and grown on Boldmedia.

Each well that contained a tested coupon was inoculated with 400 μl oforganism mixture (approximately 1×10⁶ cfu/ml) making sure that the wholesurface (paint film as well as the agar surrounding it) was evenlycovered. The plates were incubated at room temp (25° C.-26° C.) withcyclic exposure to light (OTT-Lite model #OTL4012P, 40 Watt, 26 KLumen)and dark phases, for a period of four weeks. The total area covered wasevaluated at the end of each week according to percent area covered in5% increments.

Fungal efficacy was tested as follows (Modified ASTM 5590).

ASTM 5590 is a standard accelerated test method for determiningresistance of various coatings (including paints) to fungal defacement.To accommodate for high-throughput screening, this method was scaleddown from petri plates to 6-well plates. To set up the test, an agarplug was placed at the bottom of each well of the sterile 6-well plate.A single coupon was placed with a pair of sterile forceps at the centerof the agar plug (on top) with the painted surface facing upwards.Fungal inoculums were prepared by mixing equal concentrations(approximately 1×10⁶ cfu/ml) and equal volumes (depending on number ofsamples to be tested) of like growing organisms. For this synergy study,three pools of mixed fungi were prepared as the test inoculum.Cladosporium herbarum was mixed with Trichoderma viride, Aspergillusniger was mixed with Penicillium funiculosum and Alternaria alternatawas mixed with Aureobasidium pullulans. Each well was inoculated with400 microliter of organism mixture (approximately 1×10⁶ cfu/ml) makingsure that the whole surface (paint film as well as the agar surroundingit) was evenly covered. The plates were incubated at 30° C. in presenceof moisture, for a period of four weeks. The total percent area coveredwas evaluated and recorded at the end of each week and recorded inincrements of 5%.

The Synergy Index calculation was performed as follows.

The SI is calculated based on F. C. Kull et. Al. method (AppliedMicrobiology, Vol. 9 (1961). In this study, SI was calculated based onthe following formula with the minimum inhibitory concentration chosenbased on the percent inhibitory exhibited by the individual biocideagainst each microorganisms tested.

SI=Qa/QA+Qb/QB

-   -   Qa=the concentration of Biocide A in the blend    -   QA=The concentration of Biocide A as the only biocide    -   Qb=The concentration of Biocide B in the blend    -   QB=The concentration of Biocide B as the only biocide

SI value of <1 in the formula indicates a synergism of the blendedbiocides exists.

Note: If any of the active with maximum concentration tested did notexhibit some inhibition, this maximum concentration is used to calculatethe estimated SI and a sign of less than (<) is included to take intoaccount that higher concentration of the active is needed to achieve thetargeted inhibition

NE=no end point at the concentration tested that will meet the percentinhibition criteria set in each SI calculation.

Compositions listed below that contain both zinc glyphosate and one ofDCOIT or OIT or BBIT are examples of the present invention. Othercompositions are comparative compositions.

Test results for Zinc Glyphosate and Glyphosic Acid are shown below:

% Inhibition Against Various Organisms Tested Conc. Cs + Cp + Gs + Aa +An + TV + Active ppm Ck + Nc To + Ta Os Ap Pf Ch Zinc 750 100 0 15 40 2050 Glyphosate 1500 100 0 72.5 47.5 27.5 75 2500 100 0 90 70 30 90Glyphosic Acid 750 37.5 0 15 7.5 0 2.5 1500 60 10 72.5 20 5 50 2500 807.5 90 67.5 10 72.5 Blank 100 100 100 100 100 100

Test Results for Zinc Glyphosate with DCOIT were as follows.

AaTv AnPf ApCh Cp + To + Ta Cs + Ck + Nc Gs + Os 1Zinc Glyphosate: 1DCOIT Total conc, ppm 1725 300 575 300 300 300 % inhibition 80 90 40 10080 100 SI 2.05 0.50 <0.59 0.63 0.63 0.55 1Zinc Glyphosate: 2 DCOIT Totalconc, ppm 222 222 444 222 222 444 % inhibition 60.0 80 70 100 80 70 SI0.26 0.31 <0.55 0.44 0.44 0.71 1Zinc Glyphosate: 5 DCOIT Total conc, ppm690 690 690 690 690 1035 % inhibition 80 80 70 80 100 60 SI 0.81 0.78<1.03 1.27 1.28 1.42 1Zinc Glyphosate: 10 DCOIT Total conc, ppm 632.5632.5 632.5 949 949 949 % inhibition 60 90 90 80 60 70 SI 0.74 0.64<1.02 1.71 1.71 1.21 3Zinc Glyphosate: 1 DCOIT Total conc, ppm 412 412825 412 412 412 % inhibition 70 80 90 80 100 100 SI 0.49 0.84 <0.27 0.930.93 0.87 5Zinc Glyphosate: 1 DCOIT Total conc, ppm 384 825 1650 384 384384 % inhibition 80 90 70 80 100 100 SI 0.46 1.79 <0.89 0.88 0.88 0.8410Zinc Glyphosate: 1 DCOIT Total conc, ppm 825 825 825 825 825 825 %inhibition 80 80 70 100 100 100 SI 1.00 1.88 <0.36 1.95 1.95 1.91 ZincGlyphosate Total conc, ppm 825 412.5 3300 412.5 412.5 412.5 % inhibition40 80 0 80 80 80 DCOIT Total conc, ppm 287.5 1150 575 575 575 825.5 %inhibition 40 80 40 80 20 60 Zinc Glyphosate + DCOIT at ratio 1:10 to10:1 exhibited a synergy. Zinc Glyphosate + DCOIT at ratio 0.33:1 to 2:1showed broad spectrum synergy.

Test Results for Zinc Glyphosate with OIT were as follows:

AaTv AnPf ApCh Cp + To + Ta Cs + Ck + Nc Gs + Os 1Zinc Glyph:1 OIT Totalconc, ppm 3300 3300 NE 1650 2475 825 % inhibition 40 40 80 85 80 SI<4.17 4.17 2.61 4.38 0.96 1Zinc Glyph:2 OIT Total conc, ppm 2700 6751350 1350 2025 1350 % inhibition 60.0 60 40 80 80 80 SI <4.27 1.07 <1.241.76 3.14 1.55 1Zinc Glyphosate:5 OIT Total conc, ppm 2160 2160 NE 5402160 2160 % inhibition 60 40 80 100 40 SI <4.11 4.11 0.55 2.87 2.441Zinc Glyphosate:10 OIT Total conc, ppm 1980 1485 NE 1980 1980 NE %inhibition 50 40 100 100 SI <4.05 3.04 1.77 2.44 3Zinc Glyphosate:1 OITTotal conc, ppm 825 825 NE 3300 412 412 % inhibition 55 40 60 85 80 SI<0.65 0.65 6.61 0.86 0.49 5Zinc Glyphosate:1 OIT Total conc, ppm 33001650 NE 3300 1650 825 % inhibition 50 40 70 85 90 SI <2.06 1.03 7.073.64 0.99 10Zinc Glyphosate:1 OIT Total conc, ppm NE 3300 NE 3300 1650825 % inhibition 50 80 70 80 SI 1.58 7.49 3.80 0.99 Zinc GlyphosateTotal conc, ppm 3300 1650 3300 412.5 412.5 825 % inhibition 0 40 0 80 8080 DCOIT Total conc, ppm 450 450 900 1350 900 900 % inhibition 100 50 4080 100 80

Test Results for Zinc Glyphosate with BBIT were as follows:

Cp + To + Ta + Cs + Ck + Nc 3ZnGlyphosate:1BBIT Total conc in pppm 2475% Inhibition 0 SI <1.0 1ZnGlyphosate:10BBIT Total conc in pppm 1485 %Inhibition 0 SI <1.04 1ZnGlyphosate:10BBIT Total conc in pppm 990 %Inhibition 0 SI <0.69 1ZnGlyphosate:5BBIT Total conc in pppm 1620 %Inhibition 0 SI <0.69 1ZnGlyphosate:1BBIT Total conc in pppm 1650 %Inhibition 0 SI <0.86 ZnGlyphosate Total conc in pppm 3300 % Inhibition100, NE BBIT Total conc in pppm 1350 % Inhibition 0Zinc Glyphosate+BBIT at ratio 1:1 to 3:1 and 1:5 and 1:10 exhibited asynergy. Three more separate studies were conducted with the mixtures ofalgae: Gs+Os, fast growing fungi: An, Aa, Pf, Tv and slow growing fungi:Ap+Ch. No synergistic effect was observed from those three studies.

1. A synergistic antimicrobial composition comprising a glyphosatecompound and one of 4,5-dichloro-2-n-octyl-4-isothiazolin-3-one or2-n-octly-4-isothiazolin-3-one or N-Butyl-1,2-benzisothiazolin-3-one. 2.The synergistic antimicrobial composition of claim 1, wherein saidglyphosate compound comprises zinc glyphosate.
 3. The synergisticantimicrobial composition of claim 2, wherein said synergisticantimicrobial composition comprises4,5-dichloro-2-n-octyl-4-isothiazolin-3-one, and wherein the weightratio of said 4,5-dichloro-2-n-octyl-4-isothiazolin-3-one to said zincglyphosate is from 0.1:1 to 10:1.
 4. The synergistic antimicrobialcomposition of claim 3, wherein said weight ratio of said4,5-dichloro-2-n-octyl-4-isothiazolin-3-one to said zinc glyphosate isfrom 0.33:1 to 2:1.
 5. The synergistic antimicrobial composition ofclaim 2, wherein said synergistic antimicrobial composition comprises2-n-octyl-4-isothiazolin-3-one, and wherein the weight ratio of said2-n-octyl-4-isothiazolin-3-one to said zinc glyphosate is from 0.1:1 to10:1.
 6. The synergistic antimicrobial composition of claim 5, whereinsaid weight ratio of said 4,5-dichloro-2-n-octyl-4-isothiazolin-3-one tosaid zinc glyphosate is from 0.3:1 to 0.4:1.
 7. The synergisticantimicrobial composition of claim 2, wherein said synergisticantimicrobial composition comprises N-Butyl-1,2-benzisothiazolin-3-one,and wherein the weight ratio of said N-Butyl-1,2-benzisothiazolin-3-oneto said zinc glyphosate is from 0.1:1 to 10:1.
 8. A method of inhibitingthe growth of or controlling the growth of microorganisms in a buildingmaterial, said method comprising the step of adding the synergisticantimicrobial composition of claim 2 to said building material.
 9. Acoating composition comprising the synergistic antimicrobial compositionof claim
 2. 10. A dry film made by a process comprising applying a layerof the coating composition of claim 9 to a substrate and drying saidcoating composition or allowing said coating composition to dry.